OMIPs Directory

OMIP-063

28‐Color Flow Cytometry Panel for Broad Human Immunophenotyping

Cytometry PART A, Volume 97, Issue 8, 777-781 (2020)
Kathryn Payne, Wenyan Li, Robert Salomon, Cindy S. Ma

Description: A 28-color panel was developed to screen for a range of lymphocyte subsets in human peripheral blood mononuclear cells (PBMCs), particularly in patients with primary immunodeficiency (PID). Using the panel, we are able to avoid running the sample over multiple screening panels while still deeply phenotyping a diverse range of lymphocyte subsets including innate like lymphocytes (γδ), mucosal-associated invariant T (MAIT), natural killer (NK), and NKT cells, as well as multiple subsets of naïve and memory CD4+ and CD8+ T cells, and B cells. Specifically, naïve (tn), central memory (Tcm) and effector memory (Tem) CD4+ T cells, naïve Tcm, Tem, and CD45RA+ revertant memory (TEMRA) CD8+ T cells, regulatory (Tregs), T follicular helper (Tfh) and T helper (Th) 1, and Th17 CD4+ T cells, and transitional, naïve, memory, and CD21+ B cells.

Species: Human

Tissue: Peripheral blood

Cell Subset: B cell, B memory, Transitional b, Mait, T memory, Tcrgd, Th1, Th17, Treg, Nk, Nkt, Tfh

Keyword: Primary immunodeficiency (pid)

Machine: BD FACSymphony A5

Panel details

Marker Species Clone Fluorophore Purpose Co-expressions groups Antigen Density
CD185 Human RF8B2 Brilliant Ultraviolet 615 Tfh B cell, T helper, B memory, TCRgd Low
CD19 Human SJ25C1 Brilliant Violet 750 B Cell B cell, B memory Low
CD21 Human B-ly4 Brilliant Ultraviolet 563 B Cell (Down-regulated on atypical/aged memory B cells) B cell, B memory Low
IgG Human G18-145 Brilliant Blue 660-P2 IgG+ Isotype switched memory B cells B memory High
CCR6 Human 11A9 Brilliant Blue 630-P2 Th17 T cell, T helper, TCRgd Med
IgM Human G20-127 APC-R700 Naïve B cells and IgM+ memory B cells B cell, B memory Med
CD25 Human 2A3 cFluor YG584 Treg T cell, Treg Low
Dead cells Human Zombie UV Viability
CD20 Human 2H7 Brilliant Ultraviolet 805 B Cell B cell, B memory Med
CD4 Human SK3 Brilliant Ultraviolet 661 T helper T cell, T helper, T memory High
CD161 Human DX12 Brilliant Violet 786 MAIT, NK, NKT, and a subset of CD8+ T cells MAIT, NKT Low
TCR gamma/delta Human 11F2 Brilliant Violet 711 γδ T cell TCRgd Low
CD10 Human HI10a Brilliant Violet 650 Transitional B cells B cell Low
CD3 Human UCHT1 Brilliant Violet 570 T cell T cell, T helper, T memory High
IgD Human IA6-2 Brilliant Violet 480 Naive B cells, low levels on IgM memory B cells B cell, B memory High
CD183 Human 1C6 Brilliant Violet 421 Th1 T cell, T helper Med
CD127 Human hIL-7R-M21 Brilliant Blue 700 Down-regulated on Tregs T cell, Treg Low
CD8a Human RPA-T8 Brilliant Ultraviolet 496 Cytotoxic T cell T cell, T memory High
CCR7 Human G043H7 PE-Cy7 Naïve T and CMEM T cells T cell, T memory Low
KLRG1 Human 2F1/KLRG1 APC senescence on CD8+ T cells T cell Low
CD56 Human NCAM16.2 Brilliant Blue 790-P NK cells and some memory CD8+ T cells NK Low
TCR alpha/beta Human T10B9.1A-31 Brilliant Ultraviolet 737 T cell (conventional aß T) T cell, T helper, T memory Med
CD45RA Human HI100 Brilliant Ultraviolet 395 naïve and revertant effector (TEMRA) T cells T cell, T memory Med
CD27 Human M-T271 Brilliant Blue 515 Memory B cells; naïve and cmem CD4+ and CD8+ T cells T cell, TCRgd, T memory Med
PD-1 Human EH12.1 PE-CF594 Activation T cell Low
TCR V alpha 7.2 Human 3C10 APC-Cy7 T cell receptor for MAIT cells MAIT Med
IgA1/IgA2 Human G20-359 PE-Cy5 IgA+ Isotype switched memory B cells B cell High